New primers for fast detection of Giardia duodenalis assemblages A and B using realtime PCR

Piotr Solarczyk,

Agnieszka Wojtkowiak-Giera,

Marcin Hołysz,

Anna Słodkowicz-Kowalska,

Paweł P. Jagodziński,

Krzysztof Stojecki,

Anna Rocka,

Anna C. Majewska,

Łukasz Skrzypczak

Abstrakt

Abstract. Giardia duodenalisis one of the six Giardia species and itis the most common, cosmopolitan flagellate that infects humans and many species of animals.This species exhibits considerable genetic diversity; to date, eight assemblages (A–H) have been defined. These assemblages differ in host specificity: assemblages A and B have beenfound in both humans and in many animal species. Mixed infections with Giardia (A and B) assemblages have been reported in humans and in animals. Many molecular techniques are effective and rapid for the detection of G. duodenalis and also forthe determination of genetic variability of isolates in clinical and environmental samples. In this context, the aim of this study was to design new assemblage-specific primers for rapid detection and identification ofG. duodenalis assemblages A and B and both of these assemblages simultaneously using quantitative real-time polymerase chain reaction (qPCR). Fragments of glutamate dehydrogenase and triose phosphate isomerase were used as targets in the design of primers.
In conclusion, the use of G. duodenalis assemblage-specific primers designed in this study allows quick identification of human infectious G. duodenalis assemblages A and B as well as mixed AB assemblages in a sample without further sequencing of the amplification products, which reduces the cost of study and the waiting time for the results.